Methods and compositions to reduce fat gain, promote weight loss in mammals

ABSTRACT

The present disclosure relates to compositions and related methods to reduce body weight, body fat, waist and hip size, plasma total cholesterol, LDL, triglycerides, blood glucose, leptin and C-reactive protein levels and increase in HDL and serotonin levels in an mammal. The LeptiCore® formulation disclosed, at both the low and high dosages, is helpful in the management of fat gain and its related complications. The higher dosage resulted in significantly greater reductions in body weight and triglyceride, blood glucose, and C-reactive protein levels, as well as increased serotonin levels.

This application claims priority benefit to U.S. Provisional ApplicationNo. 61/317,660, filed on Mar. 25, 2010, the contents of which areincorporated by reference herein in its entirety.

BACKGROUND

1. Field

This disclosure generally relates to compositions and methods thatimprove various health related factors and conditions. Morespecifically, the present disclosure relates to methods and compositionscomprising LeptiCore®, which is a combination of various ingredients, topromote weight loss in obese and overweight human subjects and improvevarious health related parameters associated with obesity and metabolicsyndrome. Additionally, the present disclosure also relates tocompositions and related methods to reduce body weight, body fat, waistand hip size, plasma total cholesterol, LDL, triglycerides, bloodglucose, leptin and C-reactive protein levels and increase in HDL andserotonin levels in a mammal.

2. General Background

Metabolic syndrome, a common disorder also known as Syndrome X andinsulin resistance syndrome is (like many other obesity-relatedconditions) on the rise throughout the world. The American HeartAssociation criteria for metabolic syndrome include abdominal obesity,atherogenic dyslipidemia, elevated blood pressure, insulin resistance orglucose intolerance, prothrombotic state, and pro-inflammatory state.

As of 2001, some 47 million people in the United States had metabolicsyndrome—which was projected to overtake cigarette smoking as thegreatest risk factor of cardiovascular disease [1]. One of theaccompanying conditions of metabolic syndrome that has not received asmuch attention is oxidative stress. This seems to be a key in thedisease condition, and all efforts to address the independent componentsof metabolic syndrome should include an effort to reduce oxidativestress, which has not been clearly established to be a prelude orconsequence of metabolic syndrome.

In an effort to delay the onset of heart disease and type-2 diabetes forthe vast and ever-increasing numbers of overweight/obese persons now athigh risk for these diseases, the current International DiabetesFoundation recommends pharmacological agents that specifically targetindividual components of metabolic syndrome [2-7]. Although these agentscan have benefits, the combination of safety concerns, high costs, andthe complex nature of the condition have encouraged the investigation ofmedicinal plants as natural supplements in its treatment.

LeptiCore® is a blend of plant-based polysaccharides, esterified fattyacids, pomegranate, polyphenols and ellagic acid, beta-carotene, andaphanizomenon flosaquae extract, which are components shown to reducestored body fat, enhance weight loss, act as antioxidants and amelioratethe symptoms of metabolic syndrome. The exact formulation of LeptiCore®is shown in FIG. 1.

Research has shown that leptin serves as a regulator of body fat storageby modulating satiation, glycemic control and metabolism; and thatreduction in serum leptin correlates with lower regional body fat andtotal body fat [8-10]. Leptin has also been shown to cause oxidativestress [11], which can further exacerbate metabolic syndrome.

Leptin is secreted by fat—the more fat, the more leptin—yet it is namedfor the Greek word leptos, which means “thin.” While obese peopleproduce much higher levels of leptin than thin and normal-weightindividuals, they are somehow resistant to its effects [8, 12].

One reason for this may be that elevated levels of C-reactive protein(CRP), which inhibits leptin's role in controlling appetite [13]. CRP isnow considered a key marker for low-grade systemic inflammation which,in turn, is considered by leading scientists to be responsible for anumber of metabolic problems that result in unwanted weight gain andobesity [14]. Low grade, internal, invisible inflammation, oxidativestress, along with high leptin levels, may be a root cause of excessbody fat and the inability to lose excess or unwanted fat [15].

In vivo and in vitro research done at the University of Pittsburghdemonstrated that CRP binds to leptin thereby impairing its signaling intwo ways: 1) the coupling of CRP to leptin makes crossing theblood-brain barrier nearly impossible, thus preventing “free” leptinaccess to the hypothalamus; 2) the CRP/leptin complex inhibits thebinding of leptin to its receptors, thus blocking its ability to signalin cultured cells. Since human CRP correlates with increased plasmaleptin and adiposity, the results “suggest a potential mechanism toleptin resistance, by which circulating CRP binds to leptin andattenuates its physiological functions [16].

In one set of studies, the researchers delivered human leptincontinuously for six days into mice which had receptors for leptin butlacked the ability to produce it. As expected, the plump mice ate lessand lost weight, and their blood glucose levels normalized. Infusionscontaining both leptin and high doses of CRP, however, blocked theaction of leptin. The plump mice just kept eating, getting fatter andfatter, and were no longer protected against diabetes. Giving CRP aloneaffected neither food intake nor body weight [16].

Another experiment showed that human liver cells increased CRPexpression when exposed to leptin, which suggests that appetite may beregulated through a feedback loop that includes the liver in addition tothe brain and leptin-secreting fat cells [16].

Of the various ingredients in the LeptiCore formulation, the plant-basedpolysaccharides and esterified fatty acids have been shown to lowerleptin level, resulting in reduced appetite and weight loss [17, 18].Various studies have shown that pomegranate extract containspolyphenolic compounds and beta-carotenes, which have considerableefficacy in lowering CRP, resulting in reduced inflammation and loweredcholesterol [19-21]. The reduction of inflammation and CRP promotesleptin's effectiveness, increasing the possibility of reduced appetiteand weight loss. These components have also been shown to be effectiveantioxidants which will consequently improve the components of metabolicsyndrome.

The last of the natural ingredients in this formulation, thephycocyanamines (blue green algae—a high phenylethylamine (PEA) blend)addresses the emotional aspects of metabolic syndrome by increasing thevital neurotransmitter, serotonin (5HT), a well known mood elevator[22,23]. Many studies have shown that serotonin affects eating behaviorand body weight, and that increased serotonin plasma levels areassociated with decreased food intake, reduced weight gain, andincreased energy expenditure [24, 25].

The primary purpose of the present study was to determine the effects oftwo dosage levels of a unique, natural formulation containing the aboveagents on weight loss in patients with metabolic syndrome.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an exemplary formulation, as described and claimed in atleast one embodiment.

SUMMARY

Methods

The study was an 8 week randomized, double blind, placebo-controlleddesign involving 92 obese (mean 8MI>30 kg/m2) participants (37 males; 55females; ages 1952; mean age=30.7). The participants were randomlydivided into three groups: placebo (n=30), LeptiCore® formula A (IOWdose) (n=31) and LeptiCore® formula B (high dose) (n=31). Capsulescontaining the placebo or active formulations were administered twicedaily before meals with 300 ml of water. None of the participantsfollowed any specific diet nor took any weight-reducing medications forthe duration of the study. A total of 12 anthropomorphic and serologicalmeasurements were taken at the beginning of the study and after 2, 4, 6,and 8 weeks of treatment.

Results

Compared to the placebo group, the two active groups showedstatistically significant differences on all 12 variables by week 8.These included four anthropomorphic variables (body weight, body fat,waist and hip size) and eight measures of serological levels (plasmatotal cholesterol, LDL, HDL, triglycerides, blood glucose, serotonin,leptin, C-reactive protein). The two active groups also showedsignificant intra-group differences on all 12 variables between studyonset and week 8.

DETAILED DESCRIPTION

Participants

Ninety-two obese participants were recruited for the 8-week study. Basedon physical examination by a physician and laboratory screening tests,all participants met the American Heart Association criteria formetabolic syndrome, which include abdominal obesity, atherogenicdyslipidemia, elevated blood pressure, insulin resistance or glucoseintolerance, prothrombotic state, and pro-inflammatory state.

Specific inclusion criteria included: BMI>30 kg/m2; totalcholesterol>200 mg/dl; LDL cholesterol>160 mg/dl; HDL cholesterol<40mg/dl; triglycerides>150 mg/dl; fasting blood glucose>100 mg/dl; andblood pressure>130/85 mmHg.

Specific exclusion criteria included: morbid obesity (BMI>40 kg/m2);diabetes mellitus requiring daily insulin management;pregnancy/lactation; active infection; and systemic disease such asHIV/AIDS, active hepatitis or clinical signs of active malignancy withinthe past 5 years. Other exclusion criteria included the use of anycholesterol-lowering medications 30 days prior to enrolling in the studyand/or medication (e.g., steroids) that interfere with healing; andenrollment in another clinical study within the past 6 months.

Of the 92 subjects, 37 were male and 55 were female. The age range was19-52 (mean age=30.7), and the mean BMI was >30 kg/m2.

The purpose, nature and potential risks of the study were explained toall patients and all gave a written informed consent beforeparticipation. The Cameroon National Ethics Committee approved theprotocol. The study was conducted in accordance with the HelsinkiDeclaration (1983 version). None of the participants took anyweight-reducing medication nor followed any specific diet during theduration of the study.

Study Design/Intervention

The study was a randomized, double blind, placebo-controlled design.Participants were randomly divided into three groups: placebo,LeptiCore® formula A (low dose) and LeptiCore® formula B (high dose).The placebo or active formulations were administered twice daily beforemeals with approximately 300 ml (8-10 oz.) of water. Formula A=1 capsule(300 mg each); Formula B=2 capsules (300 mg each). Since the active andplacebo capsules were identical in shape, color, and appearance, neitherthe participants nor the researchers knew which capsule wasadministered.

All product formulations were supplied by Pipeline Nutraceuticals, Inc.Fairfield, Calif., USA. All encapsulating, bottling and packaging wassupplied by and carried out by Protein Research, Inc., Livermore,Calif., USA.

During the study period, subjects were examined bi-weekly. Their bodyweight, percent body fat, and waist/hip circumferences were recorded,and serological analysis was performed. Subjective impressions (e.g.,increased/decreased appetite, feelings of lightness, gastrointestinalpains, etc.) were solicited and recorded during each visit. The subjectswere also asked about their physical activity and food intake althoughno major dietary changes or exercises were suggested.

Anthropometric Measurements

Body weight, body fat, waist and hip circumference were assessed at eachvisit with a Tanita™ BC-418 Segmental Body Composition Analyzer/Scalethat uses bioelectrical impedance analysis for body compositionanalysis. Height was measured with a Harpended™ stadiometer, whichmeasures the length of curved line staffage to the nearest 0.5 cm.

Participants (12-hour fasted) were encouraged to wear light clothingbefore measurements were taken. Waist and hip measures were obtainedwith a soft non-stretchable plastic tape. Waist circumference wasmeasured on the narrowest and the widest parts of the trunk; hipcircumference was obtained at the widest point of the hip. In an effortto ensure intra-individual consistency, the participants were measuredat approximately the same time of day each visit.

Serological/Laboratory Methods

Blood samples were collected after a 12-hour overnight fast intoheparinized tubes at the beginning of the study and after 2, 4, 6, and 8weeks of treatment. The concentrations of total cholesterol, LDLcholesterol, HDL cholesterol, triglycerides, fasting blood glucose,serotonin, C-reactive protein, and leptin were measured using commercialdiagnostic kits from SIGMA Diagnostics, St. Louis, Mo., USA.

Statistical Analysis

The data for each parameter were summarized (n, mean, standarddeviation) for Week 0 (Initial) and Week 8 (final) and for theintra-group percent differences. The percent change from baseline wastested for differences using the Mixed Effects Model, a flexible toolfor analyzing repeated and longitudinal treatments.

Results

Anthropomorphic characteristics (body weight, body fat, waist size, hipsize)

As shown in Tables 1, 2, 3, and 4, over a period of 8 weeks the twotreatment groups (Formula A=low dose; Formula B=high dose) showed astatistically significant decrease compared to the placebo group on allfour variables. The only significant difference between the twoformulations was in body weight loss (see Table 1).

Although the placebo group showed virtually no change (0.82 kg) in bodyweight over the 8-week trial, the Formula A group lost 5.2 kg (p<0.001),and the Formula B group lost 6.6 kg (p<0.001). In terms of intra-groupmean-% change from baseline (Initial) to 8 weeks, the placebo, Formula Aand Formula B groups lost 0.82%, 5.4% (p<0.05), and 6.95% (p<0.001),respectively.

TABLE 1 Body weight: effectiveness of treatments Weight Body Weight(mean kg) Change (%) Initial 8 Weeks 8 Weeks − Initial Formula B 100.93± 6.30  94.36 ± 5.67^(b)* −6.95^(‡) (↑ dose) Formula A 102.28 ± 7.17 97.04 ± 5.95^(b) −5.40^(†) (↓ dose) Placebo 101.32 ± 6.13 100.50 ± 7.28−0.82 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05; **p <0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 compared withInitial; intragroup analysis Kuate et al. Lipids in Health and Disease2010 9:20 doi: 10.1186/1476-511X-9-20

As with body weight, the placebo group showed little (0.1%) change inmean %-body fat after 8 weeks whereas the Formula A group lost 1.1%(p<0.05), and the Formula B group lost 1.8% (p<0.05). In terms ofintra-group mean % change from baseline to 8 weeks, the placebo, FormulaA and Formula B groups lost 0.24%, 2.8% (p<0.05), and 4.1% (p<0.001),respectively.

TABLE 2 Body fat: effectiveness of treatments Body Fat (mean %) FatReduction (%) Initial 8 Weeks 8 Weeks − Initial Formula B (↑ dose) 43.85± 5.60 42.04 ± 6.63^(a) −4.13^(‡) Formula A (↓ dose) 40.22 ± 7.26 39.08± 6.12^(a) −2.83^(†) Placebo 42.28 ± 8.27 42.18 ± 8.117 −0.24 ^(a)p <0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05; **p < 0.001compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 compared withInitial; intragroup analysis Kuate et al. Lipids in Health and Disease2010 9:20 doi: 10.1186/1476-511X-9-20

Waist circumference is one of the most important determinants in thediagnosis of obesity and metabolic syndrome. In this case, the Formula Aand Formula B groups showed a 4.6 cm and 5.6 cm decrease, respectively,vs. the 0.6 cm decrease demonstrated by the placebo group (p<0.05). Interms of intra-group mean % change in waist size from baseline to 8weeks, the placebo, Formula A and Formula B groups lost 0.57%, 4.4%(p<0.05), and 4.6% (p<0.001), respectively.

TABLE 3 Waist size: effectiveness of treatments Waist Circumference(mean cm) Waist Change (%) Initial 8 Weeks 8 Weeks − Initial Formula B110.50 ± 7.28 105.38 ± 10.64^(a) −4.64^(‡) (↑ dose) Formula A 104.00 ±8.63  99.40 ± 11.73^(a) −4.42^(†) (↓ dose) Placebo 105.60 ± 7.91 105.00± 15.77 −0.57 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p <0.05; **p < 0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001compared with Initial; intragroup analysis Kuate et al. Lipids in Healthand Disease 2010 9:20 doi: 10.1186/1476-511X-9-20

Once again, although the placebo group showed virtually no change (0.4cm), the Formula A group lost 4.4 cm (p<0.05), and the Formula B grouplost 5.6 cm (p<0.05) in circumference over the 8-week trial. In terms ofintra-group mean % change from baseline to 8 weeks, the placebo, FormulaA and Formula B groups lost 0.32%, 3.5% (p<0.05), and 4.6% (p<0.001),respectively.

TABLE 4 Hip size: effectiveness of treatments Hip circumference (meancm) Hip change (%) Initial 8 Weeks 8 Weeks − Initial Formula B 123.38 ±10.84 117.75 ± 17.24^(a) −4.56^(‡) (↑ dose) Formula A 126.20 ± 12.91121.80 ± 15.41^(a) −3.49^(†) (↓ dose) Placebo 125.85 ± 14.72 125.45 ±17.99 −0.32 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05;**p < 0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 comparedwith Initial; intragroup analysis Kuate et al. Lipids in Health andDisease 2010 9:20 doi: 10.1186/1476-511X-9-20

Serological characteristics (total cholesterol, LDL cholesterol, HDLcholesterol, triglycerides, fasting blood glucose, serotonin, leptin,C-reactive protein)

As shown in Tables 5, 6, 7, 8, 9 and 10, the two treatments (vs.placebo) groups showed a statistically significant difference on alleight variables by the end of the 8-week trial period. There weresignificant differences between the two formulations (in favor of thehigh dosage Formula B) on triglyceride, blood glucose, serotonin, andC-reactive protein levels.

In contrast to the small (4.5 mg/dL) reduction in total cholesterolshown by the placebo group at 8 weeks, the reduction for the Formula Agroup was 37.6 mg/dL (p<0.05), and that of the Formula B group was 57.9mg/dL (p<0.001). In terms of intra-group mean % change from baseline(Initial) to 8 weeks, the placebo group decrease was 2.1% whereas theFormula A and Formula B groups decreases were 18.0% (p<0.05) and 27.5(p<0.001), respectively.

As with total cholesterol level, the placebo group showed a small (3.6mg/dL) decrease in LDL cholesterol by 8 weeks. This can be compared withthe 24.0 mg/dL reduction shown by the Formula A group (p<0.05) and the32.8 mg/dL reduction of the Formula B group (p<0.001). In terms ofintra-group mean % change in LDL from baseline to 8 weeks, the placebo,Formula A and Formula B groups decreased by 2.4%, 14.7% (p<0.05), and19.8% (p<0.001), respectively.

On one of the few variables where an increase in level is desirable, theplacebo group showed a very small (1.3 mg/dL) increase in HDLcholesterol at 8 weeks. This can be compared with the 4.7 mg/dL increaseshown by the Formula A group (p<0.05) and the 6.2 mg/dL increase of theFormula B group (p<0.001). In terms of intra-group mean % change in HDLfrom baseline to 8 weeks, the placebo, Formula A and Formula B groupsincreased by 3.6%, 13.6% (p<0.05), and 19.1% (p<0.001), respectively.

TABLE 5 Plasma Total Cholesterol, LDL cholesterol and HDL cholesterollevels: effectiveness of treatments Time (weeks) % Change Variable GroupInitial 8 8 Weeks − Initial Total Cholesterol Placebo 211.16 ± 11.92206.68 ± 20.19 −2.12 (mg/dl) Formula A (↓dose) 208.74 ± 10.49 171.17 ±18.82^(a) −18.00^(†) Formula B (↑dose) 210.64 ± 9.30  152.75 ± 10.19^(b)−27.48^(‡) LDL Cholesterol Placebo 166.21 ± 9.25  162.63 ± 17.83 −2.39(mg/dl) Formula A (↓dose) 163.94 ± 8.27  139.91 ± 8.92^(a) −14.66^(†)Formula B (↑dose) 165.84 ± 7.29  133.05 ± 7.71^(b) −19.77^(‡) HDLCholesterol Placebo 35.92 ± 5.16  37.22 ± 5.59 +3.62 (mg/dl) Formula A(↓dose) 34.72 ± 3.28  39.46 ± 4.48^(a) +13.65^(†) Formula B (↑dose)32.43 ± 2.10  38.63 ± 3.11^(b) +19.12^(‡) ^(a)p < 0.05; ^(b)p < 0.001compared with Placebo *p < 0.05; **p < 0.001 compared with Formula A^(†)p < 0.05; ^(‡)p < 0.001 compared with Initial; intragroup analysisKuate et al. Lipids in Health and Disease 2010 9:20 doi:10.1186/1476-511X-9-20

Per the above, the placebo group demonstrated a small change (1.0 mg/dLdecrease) in triglyceride level at the end of the 8-week trial. This canbe compared with the 11.5 mg/dL decrease shown by the Formula A group(p<0.05) and the 27.6 mg/dL reduction demonstrated by the Formula Bgroup (p<0.001). In terms of intra-group mean % change in triglyceridelevel from baseline to 8 weeks, the placebo, Formula A and Formula Bgroups decreased by 0.6%, 7.1% (p<0.05), and 16.4% (p<0.001),respectively.

TABLE 6 Plasma Triglyceride level: effectiveness of treatmentsTriglyceride change (%) (mean mg/dl) 8 Weeks − Initial 8 Weeks InitialFormula B (↑ dose) 167.61 ± 10.84 140.04 ± 4.29^(b)* −16.45^(‡) FormulaA (↓ dose) 162.21 ± 12.95 150.71 ± 8.30^(a) −7.09^(†) Placebo 168.93 ±14.04 167.90 ± 10.30 −0.61 ^(a)p < 0.05; ^(b)p < 0.001 compared withPlacebo *p < 0.05; **p < 0.001 compared with Formula A ^(†)p < 0.05;^(‡)p < 0.001 compared with Initial; intragroup analysis Kuate et al.Lipids in Health and Disease 2010 9:20 doi: 10.1186/1476-511X-9-20

In contrast to the small (1.6 mg/dL) decrease in blood glucose levelshown by the placebo group at 8 weeks, the reduction for the Formula Agroup was 8.4 mg/dL (p<0.05) and that of the Formula B group was 12.8mg/dL (p<0.001). In terms of intra-group mean % change from baseline to8 weeks, the placebo, Formula A and Formula B groups decreased by 1.5%,7.9% (p<0.05), and 12.9% (p<0.001), respectively.

TABLE 7 Fasting Blood Glucose levels: effectiveness of treatments BloodGlucose (mean mg/dl) change (%) Initial 8 Weeks 8 Weeks − InitialFormula B 104.84 ± 9.39  92.06 ± 6.29^(b)* −12.19^(‡) (↑ dose) Formula A105.61 ± 8.62  97.22 ± 7.81^(a) −7.94^(†) (↓ dose) Placebo 107.52 ± 8.73105.90 ± 10.79 −1.51 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo*p < 0.05; **p < 0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p <0.001 compared with Initial; intragroup analysis Kuate et al. Lipids inHealth and Disease 2010 9:20 doi: 10.1186/1476-511X-9-20

In another case (see HDL cholesterol above) where an increase in levelis desirable, the placebo group showed a very small (1.6 mg/dL) increasein serotonin levels at 8 weeks. This can be compared with the 9.9 mg/dLincrease shown by the Formula A group (p<0.05) and the 13.7 mg/dLincrease of the Formula B group (p<0.001). In terms of intra-group mean% change in serotonin from baseline to 8 weeks, the placebo, Formula Aand Formula B groups increased by 9.1%, 28.6% (p<0.001), and 38.6%(p<0.001), respectively.

TABLE 8 Serum Serotonin levels: effectiveness of treatments Serotonin(mean mg/dl) change (%) Initial 8 Weeks 8 Weeks − Initial Formula B35.40 ± 2.62 49.06 ± 2.85^(b)* 38.59^(‡) (↑ dose) Formula A 34.55 ± 2.8544.42 ± 2.96^(a) 28.57^(‡) (↓ dose) Placebo 34.20 ± 3.15 37.31 ± 3.699.09 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05; **p <0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 compared withInitial; intragroup analysis Kuate et al. Lipids in Health and Disease2010 9:20 doi: 10.1186/1476-511X-9-20

In contrast to the 4.4 mg/dL decrease in serum leptin levels shown bythe placebo group at 8 weeks, the reduction for the Formula A group was14.8 mg/dL (p<0.05) and that of the Formula B group was 14.38 mg/dL(p<0.001). In terms of intra-group mean % change from baseline to 8weeks, the placebo, Formula A and Formula B groups decreased by 14.4%,47.0% (p<0.001), and 48.0% (p<0.001), respectively.

TABLE 9 Serum Leptin levels: effectiveness of treatments Leptin (meanmg/dl) change (%) Initial 8 Weeks 8 Weeks − Initial Formula B (↑ dose)30.15 ± 1.50 15.83 ± 1.13^(b) −47.59^(‡) Formula A (↓ dose) 31.52 ± 1.2116.72 ± 1.40^(a) −46.95^(‡) Placebo 30.33 ± 1.63 25.97 ± 1.61 −14.38^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05; **p < 0.001compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 compared withInitial; intragroup analysis Kuate et al. Lipids in Health and Disease2010 9:20 doi: 10.1186/1476-511X-9-20

Per the above, the placebo group demonstrated a small change (0.4 mg/ldecrease) in C-reactive protein levels at the end of the 8-week trial.This can be compared with the 1.6 mg/l decrease shown by the Formula Agroup (p<0.05) and the 3.4 mg/l reduction demonstrated by the Formula Bgroup (p<0.001). In terms of intra-group mean % change in blood levelsof C-reactive protein from baseline to 8 weeks, the placebo, Formula Aand Formula B groups decreased by 3.3%, 15.1% (p<0.001), and 29.4%(p<0.001), respectively.

TABLE 10 C-Reactive Protein levels: effectiveness of treatments CRP(mean mg/l) change (%) Initial 8 Weeks 8 Weeks − Initial Formula B (↑dose) 11.58 ± 1.11  8.17 ± 0.61^(b)* −29.45^(‡) Formula A (↓ dose) 10.99± 1.26  9.33 ± 0.43^(a) −15.10^(‡) Placebo 12.46 ± 1.30 12.05 ± 1.39−3.31 ^(a)p < 0.05; ^(b)p < 0.001 compared with Placebo *p < 0.05; **p <0.001 compared with Formula A ^(†)p < 0.05; ^(‡)p < 0.001 compared withInitial; intragroup analysis Kuate et al. Lipids in Health and Disease2010 9:20 doi: 10.1186/1476-511X-9-20

Adverse Events

Adverse events with an incidence >3 included lack of sleep (4), andheadache (4). Since the type and incidence of all reported side effectswere also observed in the placebo group, one can assume that theLeptiCore® formulation had few, if any, negative side effects.

Discussion

The results herein indicate that the use of LeptiCore®, a unique naturalformulation, has considerable efficacy on weight loss in patients withmetabolic syndrome. Compared to the placebo group, the two active groups(low and high dosage) showed statistically significant differences onall 12 variables by the end of the 8-week trial. Although there were nodifferences between the two active groups on body fat, waist and hipsize; or cholesterol (total, HDL, LDL) or leptin levels, the higherdosage did result in significantly greater reductions in body weight,and triglyceride, blood glucose, CRP and serotonin levels.

It appears that LeptiCore® somehow enhances cell membrane stability forimproved cellular communication between the liver and adipose tissuesfor enhanced fatty acid utilization leading to regional fat loss. Inaddition, modulation of membrane-based inflammatory markers helped toreduce the inflammatory component of fat gain. This is where thisformulation differs from other leptin-reducing supplements. Ingredientsin this proprietary blend not only lowered serum leptin levels but alsoreduced CRP, a key marker for low-grade inflammation. As discussedearlier, some researchers believe the binding of CRP to leptin may bethe reason [16]. The fact that CRP is elevated in obese people increasesthe plausibility of their argument. CRP, which is produced by the liverand typically rises as a part of the immune system's inflammatoryresponse, has been gaining favor as a marker for hypertension and heartdisease risk—both known complications of obesity.

LeptiCore's efficacy may also be due to the fact that it effects atime-dependent transit through the gastro-intestinal tract which, inturn, elicits a signal to regulate leptin levels. This is the mechanismfor reducing the level of leptin in the bloodstream and thecorresponding reduction in the percentage of body fat. LeptiCore'sability to lower CRP levels allows the remaining circulating leptin toreside in its “free” state and bind to appetite-regulating leptinreceptors in the hypothalamus.

Yet another aspect of LeptiCore's action is its potential antioxidantpower. The plasma concentrations of leptin are markedly increased inhuman obesity and positively correlated to body fat mass [26]. As humanobesity is associated with hyperleptinemia and atherosclerosis, it wasshown that leptin, in addition to its angiogenic properties, exertsproatherogenic effect on endothelial cells by increasing reactive oxygenspecies (ROS) formation. As mentioned earlier, Boulimie et al. [11] haveshown that the administration of leptin may stimulate increases inoxidative stress in vitro cultured human endothelial cells. Increases inoxidative stress in the vascular endothelium may interact with nitricoxide to form peroxynitrite and thereby decrease the bioavailability ofnitric oxide, which, in turn, could reduce weight gain and food intake[27].

ACKNOWLEDGEMENTS

The authors are grateful to Pipeline Nutraceuticals, Inc. Fairfield,Calif., USA and Protein Research, Inc., Livermore, Calif., USA forproviding all the test material (Lepticore®) used in this study.

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While the compositions and methods have been described in terms of whatare presently considered to be the most practical and preferredembodiments, it is to be understood that the disclosure need not belimited to the disclosed embodiments. It is intended to cover variousmodifications and similar arrangements, the scope of which should beaccorded the broadest interpretation so as to encompass all suchmodifications and similar compositions and methods.

The present disclosure provides method of reducing body weight, bodyfat, waist and hip size, plasma total cholesterol, LDL, triglycerides,blood glucose, leptin and/or C-reactive protein levels and increasingHDL and/or serotonin levels in an mammal, the method comprisingproviding a composition containing one or more ingredients selected fromthe group of: Cyamopsis tetragonolobus, Guar Gum, Ceratonia siliqua,Locust Bean Gum, Amorphophallus konjac, Punica granatum—Pomegranatekonjac, Aphanizomenon Flos Aquae, Cyamopsis tetragonolobus bean,Ceratonia siliqua seed, Amorphophallus konjac root, Punica granatum seedand Aphanizomenon.

The present disclosure also provides compositions containing one or moreingredients selected from the group of: Cyamopsis tetragonolobus, GuarGum, Ceratonia siliqua, Locust Bean Gum, Amorphophallus konjac, Punicagranatum—Pomegranate konjac, Aphanizomenon Flos Aquae, Cyamopsistetragonolobus bean, Ceratonia siliqua seed, Amorphophallus konjac root,Punica granatum seed and Aphanizomenon and their related methods toreduce body weight, body fat, waist and hip size, plasma totalcholesterol, LDL, triglycerides, blood glucose, leptin and C-reactiveprotein levels and increase in HDL and serotonin levels in an mammal.

1. A composition comprising: a) Cyamopsis tetragonolobus; b) Ceratoniasiliqua; c) Amorphophallus konjac and punica granatum.
 2. Thecomposition of claim 1, wherein the composition comprises 30% cyamopsistetragonolobus, 20% ceratonia siliqua and 50% amorphophallus konjac byweight.
 3. (canceled)
 4. The composition of claim 3, wherein the punicagranatum is a seed.
 5. The composition of claim 1, further comprisingbeta-carotene.
 6. The composition of claim 5, wherein the beta-caroteneis synthetic.
 7. The composition of claim 1, further comprisingaphanizomenon flos aquae.
 8. A composition comprising: a) Cyamopsistetragonolobus; b) Ceratonia siliqua; c) Amorphophallus konjac; d)punica granatum; e) beta-carotene and f) aphanizomenon flos aquae. 9.The composition of claim 8, wherein the punica granatum is a seed. 10.The composition of claim 8, wherein the beta-carotene is synthetic.11.-20. (canceled)